Abstract
The western corn rootworm (WCR, Diabrotica virgifera virgifera) gene, dvssj1, is a putative homolog of the Drosophila melanogaster gene, snakeskin (ssk). This gene encodes a membrane protein associated with the smooth septate junction (SSJ) which is required for the proper barrier function of the epithelial lining of insect intestines. Disruption of DVSSJ integrity by RNAi technique has been shown previously to be an effective approach for corn rootworm control, by apparent suppression of production of DVSSJ1 protein leading to growth inhibition and mortality. To understand the mechanism that leads to the death of WCR larvae by dvssj1 double-stranded RNA, we examined the molecular characteristics associated with SSJ functions during larval development. Dvssj1 dsRNA diet feeding results in dose-dependent suppression of mRNA and protein; this impairs SSJ formation and barrier function of the midgut and results in larval mortality. These findings suggest that the malfunctioning of the SSJ complex in midgut triggered by dvssj1 silencing is the principal cause of WCR death. This study also illustrates that dvssj1 is a midgut-specific gene in WCR and its functions are consistent with biological functions described for ssk.
Highlights
RNA interference (RNAi) pathways are common among many eukaryotes including insects [1], and transgenic crops utilizing RNAi represent a promising new tool for insect pest management [2]
Our results show that compromised dvssj1/ssk expression caused by either RNAi- or CRISPR/Cas9-mediated systems is associated with defects in smooth septate junction (SSJ) structure, changes in the localization of certain SSJ proteins, and impaired barrier function of the midgut in both western corn rootworm (WCR) and D. melanogaster strains which culminate in impaired larval development and survival
We previously reported the discovery of the dvssj1 gene through double-stranded RNA (dsRNA) diet-based screening for targets that have the potential for WCR control [17]
Summary
RNA interference (RNAi) pathways are common among many eukaryotes including insects [1], and transgenic crops utilizing RNAi represent a promising new tool for insect pest management [2]. The western corn rootworm (WCR), Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is one of the most economically important pests of maize in the United States, and Europe where it is an invasive species [3, 4]. WCR damage is managed by combinations of crop rotation, broad-spectrum soil insecticides [5], and transgenic crops expressing crystalline (Cry) proteins from Bacillus thuringiensis (Bt) [4]. Double-stranded RNA targeting the smooth septate junction of western corn rootworm to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘Author Contributions’ section
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